Identification of REV-ERB inverse agonists for cancer immunotherapy

用于癌症免疫治疗的 REV-ERB 反向激动剂的鉴定

基本信息

批准号：
10401264
负责人：
Laura A Solt
金额：
$ 61.45万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-05-01 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10401264
关键词：
Agonist Automation Binding Binding Proteins Bioinformatics Biological Assay Biological Response Modifiers Cells Collection Consensus Sequence Crystallography DNA Data Development Disease Equilibrium Exhibits Fluorescence Resonance Energy Transfer Frequencies Genes Genetic Genetic Transcription Goals Immune checkpoint inhibitor Immunologic Surveillance Immunotherapy In Vitro Inflammatory Inflammatory Bowel Diseases Interleukin-17 Lead Libraries Ligand Binding Ligands Malignant Neoplasms Mediating Multiple Sclerosis Nuclear Receptors Pharmaceutical Chemistry Pharmacology Phase I/II Clinical Trial Plasma Proteins Play Positioning Attribute Property Proteins Regimen Regulation Reproducibility Research Resistance Response Elements Role Running Series Solubility Specificity Structure-Activity Relationship T-Lymphocyte Testing Therapeutic Tissues Toxic effect Transcription Coactivator Transcription Repressor Tumor Immunity Validation anti-PD-1/PD-L1 base cancer immunotherapy chemical synthesis cheminformatics counterscreen cytokine cytotoxicity design drug discovery drug metabolism fight against follow-up genetic approach high throughput screening in vivo lead optimization meetings member miniaturize mouse model neoplasm immunotherapy novel novel strategies overexpression patient subsets pharmacophore receptor response scaffold screening small molecule therapeutic target tool transcription factor tumor

项目摘要

PROJECT SUMMARY While immunotherapy is a breakthrough in our fight against cancer, only a subset of patients display robust and long-lasting responses highlighting the critical need for novel approaches to be used alone or in combination with current therapeutic regimens. TH17 cells, which express the lineage defining transcription factor RORgt, have emerged as targets to enhance antitumor immunity, with RORg agonists currently in Phase 1/2 clinical trials. The transcriptional repressors REV-ERBa and REV-ERBb are often co-expressed in the same tissues as RORg(t) and bind the same DNA response elements, which suggests mutual cross talk and co-regulation of their target genes. Our preliminary data demonstrates that the REV-ERBs are ligand-regulated transcription factors and play critical roles in TH17 responses, including regulation of IL-17A expression through competition with RORgt at their shared DNA consensus sequence(s), and regulation of RORgt expression itself. We have generated small molecule ligands that enhance the REV-ERBs repressive function and inhibit TH17 cell development in vitro and in vivo. Thus, we hypothesize that identification of ligands that inhibit the REV-ERBs repressive activity will enhance TH17 responses and antitumor immunity. In order to identify ligands that inhibit REV-ERB activity, we have designed a high-throughput screening (HTS) compatible primary assay that detects direct ligand binding to each receptor. Our goal is to implement a full HTS-campaign using the Scripps Institutional Drug Discovery Library (SDDL) to identify, validate, and characterize potent small molecule inverse agonists of REV-ERB activity. To achieve our goal, we will miniaturize our assay into a 1,536-well plate format and upon meeting HTS automation criteria, carry out a “full-deck” HTS campaign to screen the >640,000 compounds in the SDDL (Aim 1). Cheminformatic analysis of “hits” will help identify the most promising leads by structural clustering, bioinformatics analysis of compounds to determine promiscuity, and scaffold analysis to determine ease of chemical synthesis and tractability for further medicinal chemistry efforts to perform structure- activity relationship studies. In Aim 2, we will use a cascade of follow up assays to validate screening hits, determine specificity, and begin to understand mechanism of action of REV-ERB-mediated transcriptional activity. Finally, in Aim 3, validated HTS hits will be advanced to early medicinal chemistry for lead optimization and characterization of novel negative regulators of REV-ERBa/b. We expect that completion of this application will deliver multiple structurally distinct REV-ERB inverse agonists that exhibit suitable levels of cellular activity, potency, and selectivity. Our collaborative research team has a strong track record of performing high-throughput screens, selective optimization of scaffolds, and in vitro and in vivo characterization of compounds. Collectively, our screening approach puts us in a unique position to identify, validate, and characterize novel REV-ERB- selective small molecule inverse agonists for the study of the REV-ERB’s function in enhancing TH17 responses and to determine whether targeting these receptors is a viable option for immunotherapy.

项目概要虽然免疫疗法是我们抗击癌症的一项突破，但只有一小部分患者表现出稳健的表现持久的反应强调了单独或组合使用新方法的迫切需要使用当前的治疗方案，TH17细胞表达谱系定义的转录因子RORgt，已成为增强抗肿瘤免疫的靶标，RORg激动剂目前处于1/2期临床转录抑制因子 REV-ERBa 和 REV-ERBb 通常在同一组织中共表达。作为 RORg(t) 并结合相同的 DNA 响应元件，这表明相互串扰和共同调节我们的初步数据表明 REV-ERB 是配体调节的转录。因素并在 TH17 反应中发挥关键作用，包括通过竞争调节 IL-17A 表达与 RORgt 共享 DNA 共有序列，以及 RORgt 表达本身的调节。生成的小分子配体增强 REV-ERB 抑制功能并抑制 TH17 细胞因此，我们致力于鉴定抑制 REV-ERB 的配体。抑制活性将增强 TH17 反应和抗肿瘤免疫，以便识别抑制的配体。 REV-ERB 活性，我们设计了一种高通量筛选 (HTS) 兼容的初级检测方法，可检测 REV-ERB 活性我们的目标是使用 Scripps 实施完整的 HTS 活动。机构药物发现库 (SDDL)，用于识别、验证和表征有效的小分子逆向药物为了实现我们的目标，我们将我们的检测小型化为 1,536 孔板格式。在满足 HTS 自动化标准后，开展“全套”HTS 活动，筛选超过 640,000 SDDL 中的化合物（目标 1）对“命中”进行化学信息学分析将有助于识别最有希望的先导化合物。通过结构聚类、化合物生物信息学分析来确定混杂性，以及支架分析来确定确定化学合成的容易性和可处理性，以进行进一步的药物化学工作以进行结构- 在目标 2 中，我们将使用一系列后续测定来验证筛选命中，确定特异性，并开始了解 REV-ERB 介导的转录的作用机制最后，在目标 3 中，经过验证的 HTS 命中将被推进到早期药物化学中以进行先导化合物优化。 REV-ERBa/b 的新型负调节剂的表征和表征我们预计该申请能够完成。将提供多种结构不同的 REV-ERB 反向激动剂，这些激动剂表现出适当水平的细胞活性，我们的合作研究团队在执行高通量方面拥有良好的记录。筛选、支架的选择性优化以及化合物的体外和体内表征。我们的筛选方法使我们处于独特的地位，可以识别、验证和表征新型 REV-ERB- 用于研究 REV-ERB 增强 TH17 反应功能的选择性小分子反向激动剂并确定针对这些受体是否是免疫治疗的可行选择。